International Journal of Pharmaceutical and Biological Archive

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Available Online at www.ijpscr.info
International Journal of Pharmaceutical Sciences and
Clinical Research 2021; 1(1):16-21
RESEARCH ARTICLE
Antifertility Effect of Aqueous Leaf Extract of Prosopis cineraria in Male Albino
Rats
K. Sathiyaraj1
, T. Thirumalai2
, G. Arumugam3
, Gnanavel Rathinam4
, A. Abduz Zahir5
, B. Senthilkumar6
1
SVB Foundation Trust, No.35, North Coloney, Ranipet, Tamil Nadu, India, 2
Department of Zoology,
Muthurangam Government Arts College, Vellore, Tamil Nadu, India, 3
Department of Biochemistry, Kanchi
Shri Krishna College of Arts and Science, Kanchipuram, Tamil Nadu, India, 4
Department of Botany, Sri Vijay
Vidyalaya College of Arts and Science College, Dharmapuri, Tamil Nadu, India, 5
Department of Zoology, C.
Abdul Hakeem College (Autonomous), Melvisharam, Ranipet, Tamil Nadu,India, 6
Department of Zoology,
Thiruvalluvar University, Vellore, Tamil Nadu, India
Received: 15-12-2020; Revised: 10-01-2021; Accepted: 01-02-2021
ABSTRACT
Objective: The objective of the study was to evaluate the fertility effects of aqueous leaf extract of
Prosopis cineraria treatment effects on fertility indices in male albino rats. Materials and Methods: The
aqueous leaf extract was prepared according to conventional methods. The rats were randomly divided
into three groups with six individuals in each group. Groups-I animals were used as control and given
0.5 ml distilled water by orally intubated. Group-II animals were administered 150 mg body weight of
the aqueous extract for 35 days. Group-III animals were administered 250 mg/kg body weight of the
aqueous extract for 35 days. Results: The aqueous leaf extract administered animals showed decrease
in the weight of testis, epididymis, and seminal vesicles. In addition, sperm motility, sperm viability,
and changes in testosterone levels were seen changes reproductive system, especially reduction of sperm
count, motility and abnormal head and tail condition were also observed in the 250 mg/kg body weight
extract administered of the animals, when compared to the control group. Conclusion: The aqueous leaf
extract of P. cineraria showed antifertility effect in male albino rats. After the experimental period of
65 days without any oral administration of the plant extract, remarkable changes such as increase in the
weight of the testis, epididymis, and seminal vesicle and also in the increment of the sperm count were
found.
Keywords: Prosopis cineraria, Antifertility, Male Albino Rats, Testosterone, Epididymis,
Seminal vesicles
INTRODUCTION
Increase in human population throughout the
world, more particularly in developing and an
underdeveloped countries, has its detrimental
effects on the life supporting system on earth. The
possibility of an effective check on human fertility
may soon be realized through biological means.
*Corresponding Author:
Dr. K. Sathiyaraj,
E-mail: sathiyaraj.cahc@gmail.com
Fertility regulation comprising contraception and
management of infertility forms an important
component of reproductive health (Allag and
Rangari, 2002; Singh and Kala, 2011).[1-5]
Although considerable progress has been made in
the development of highly effective, acceptable,
and reversible methods of contraception among
females, progress and possibilities on males are
still sluggish and limited. With recent progress
toward a better understanding of male reproductive
physiology, there is a need to develop new

2. Sathiyaraj, et al.: Antifertility effect: An male albino rat
IJPSCR/Jan-Mar-2021/Vol 1/Issue 1 17
contraceptives. Man has been using plant materials
for medicinal purposes since time immemorial. It
may be difficult to ascertain how and where this
aspect of man-plant relationship had taken its roots,
but on the basis of empirical experience through
generations, human societies have developed a
certain system of herbal medicines.[6-12]
Herbal medicines are comparatively safer and more
hygienicthansyntheticdrugs.Plant-basedtraditional
knowledge has the distinction of being recognized
as a principal tool in the search for new sources of
drugs and pharmaceuticals (Sharma and Mujundar,
2003; Jagatheeswari, 2012). An ethnobotanical
survey can bring out many different clues for the
development of drugs to treat human diseases.
Ayurveda, the science of life, is a comprehensive
traditional system of health care in India for more
than 5000 years (Mishra et al., 2001). This system
is responsible for making preparations from Indian
medicinal plants. Ayurveda remains a vital system
of medicine and drug therapy not only in India but
also elsewhere (Borchardt, 2003). In addition to
the problem of efficacy, traditional medicine has
a problem of safety. People think that herbs are
“safe” and “harmless” since they are natural and
are not synthesized in the laboratory. However,
utilization of herbs may possibly expose the
patient to unknown dangers (Magee and Loiacono,
2004). Although traditional health systems are
locally accessible and culturally relevant, they
must first be rendered safe. Most importantly, poor
documentation, lack of standardization, and the
absence of regulatory mechanisms for traditional
health-care practice in many countries were seen as
problems to be solved (Bodeker et al., 2000).
Prosopis cineraria (Fabaceae) (Vanni Maram –
Tamil),flowerispounded,mixedwithsugar,andused
during pregnancy as safeguard against miscarriage.
The bark of the tree is dry, acrid, and bitter with a
sharptaste;coolinganthelmintic;tonic,curesleprosy,
dysentery, bronchitis, asthma, leucoderma, piles, and
tremors of the muscles. The bark is used as a remedy
for rheumatism, in cough colds, asthma. The bark is
prescribedforscorpionsting.Thesmokeoftheleaves
is good for eye problems. The fruit is dry and hot,
with a flavor, indigestible, causes biliousness, and
destroys the nails and the hair. The pod is considered
astringent in Punjab. The plant is recommended for
the treatment of snakebite (Khasgiwal et al., 1969;
Madan et al., 1972).[13-20]
MATERIALS AND METHODS
Plant material
TheleavesofP.cineraria(Fabaceae)werecollected
in and around Vellore district, Tamil Nadu. The
plant materials were cleaned with distilled water,
shade dried at room temperature, and authenticated
at the Department of Zoology, Thiruvalluvar
University, Serkadu, Vellore district, Tamil Nadu.
Plants extract preparation
The shade dried plant materials were powdered
separately in an electrical blender and stored at 5°C
untilfurtheruse.Onehundredgramsofthedriedplant
powder were taken separately mixed with 500
ml
of distilled water and then magnetically stirred in
separate containers overnight at room temperature.
The residue was removed by filtration and the
aqueous extracts were lyophilized and concentrated
under vacuum to get solid yield 10% (leaves).
Animals
Adult male Wistar albino rats weighing around
180–220 g were purchased from Tamil Nadu
VeterinaryandAnimalSciencesUniversity,Chennai,
India. The animals were kept in polypropylene
cages (four in each cage) at an ambient temperature
of 25±2°C and 55–65% relative humidity. 12±1 h
light and dark schedule was maintained in the
animal house till the animals were acclimatized
to the laboratory conditions. They were fed with
commercially available rat chow (Hindustan Lever
Ltd., Bengaluru, India) and had free access to water.
The experiments were designed and conducted in
accordance with the institutional guidelines.
Experimental Design
Dose and duration of treatment
The daily dose of the plant extract was freshly
dissolved in 0.5 mL of distilled water and orally

3. Sathiyaraj, et al.: Antifertility effect: An male albino rat
IJPSCR/Jan-Mar-2021/Vol 1/Issue 1 18
administered to each experimental animal every
morning for 35 days.
Group I: Control rats received 0.5 mL/day of the
vehicle, that is, distilled water.
Group II: Rats treated with P. cineraria at
150 mg/kg body wt. orally using intragastric tubes
for 35 days.[21-27]
Group III: Rats treated with P. cineraria at
250 mg/kg body wt. orally using intragastric tubes
for 35 days.
The plant extract was administered orally to each
treated animal every morning for 35 days. After
35 days of aqueous leaf extract treatment, half of
the animals of Groups II and III were autopsied
under light ether anesthesia. The remaining animals
were kept for recovery for a period of 65 days and
designated as recovery groups, that is, II and III.
Estimation of sperm motility and count
The spermatozoa were obtained by making
small cuts in cauda epididymis and vas deferens
and placed in 1 ml of modified Krebs-Ringer
bicarbonate buffer (pH 7.4). The sperm suspension
was evaluated for sperm count and percentage of
motility. The percentage of motility was determined
by the number progressive and non-progressive
movements of sperms observed under a compound
microscope. The sperm count was determined
under a Neubauer hemocytometer (Zaneveld, 1977;
Srikanth et al., 1999). To evaluate the abnormalities
of spermatozoa abnormalities, the sperm suspension
was stained with eosin; smears were made on slides,
air dried, and made permanent.
Serum testosterone
Serum levels of testosterone were assayed in
duplicate using specific RIA method (WHO,
Method manual 1987). Serum samples were
separated by standard procedure and stored at 20°C
for subsequent analysis.
Histological changes
The testis was removed and stored immediately
in 10% formalin initially for 48 h; thereafter, the
materials were transferred to 70% alcohol and
stored. After fixation, the tissue was subsequently
put in paraffin. Thin (5 µm) sections were
drawn using a microtome and then stained with
hematoxylin and eosin and mounted in neutral di-
styrene-dibutyl propylene medium and examined
using photomicroscopy (Strate et al., 2005).
RESULTS
Herbal medicines are popular as remedies for
diseases among a vast majority of world’s
population as remedies for diseases. From the
dawn of civilization, humans have relied on
plants and their products as a source of drugs for
their primary health care. In recent years, their
use as a popular alternative to modern medicine
has increased considerably even in developed
countries (Qureshi et al., 2006; Souad et al.,
2007). A number of plants have been identified
and evaluated by various researchers for fertility
regulation in males (Melis, 1999). In the present
study, no changes were observed in the body
weight of the treated animals. However, testicular
and epididymal weight decreased significantly
after 35 days of treatment in both 150 and 250
mg/kg body wt. treated animals. Abnormal
spermatozoa, predominantly deformed head and
tailless spermatozoa, were also observed after 35
days of treatment with higher dose regimen. The
sperm counts decreased. The sperm motility was
inhibited [Table 1]. In P. cineraria aqueous leaf
extract treated mice, the cauda epididymidal sperm
parameters showed evidence of dose-dependent
toxicity. The plant extract intoxication exerted
a significant (P 0.001) decrease in epididymal
sperm concentration, sperm progress motility, live
sperm count, and increased abnormal sperm rate.
The reduction of sperm count and sperm motility
was significantly higher in plant extract 250 mg/
kg body wt. treated animals when compared to
150 mg/kg body wt. treated and control animals.
The levels of serum testosterone in two dosage
groups were found to have significantly reduced
by when compared to the levels in control groups
(Group-I) [Table 1]. In the recovery Groups II and
III, the alteration in the concentrations of serum

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IJPSCR/Jan-Mar-2021/Vol 1/Issue 1 19
testosterone was not significant alteration when
compared to that of control animals. The light
microscopy examination of the testis of the control
rats showed normal structure completely enveloped
by a thick capsule, tunica albuginea, which is
composed mainly of dense collagenous fibrous
connective tissue. The structural components
of the testis are the seminiferous tubules and
interstitial tissues. The seminiferous tubules are
two types of cells, the Sertoli cells, resting on the
thin basal lamina (basement membrane) and the
spermatogenic cells. The testes of the plant extract
treated group (II and III) animals showed normal
features with successive stages of transformation
of the seminiferous epithelium into spermatozoa.
Leydig cells were situated in between the tubules.
DISCUSSION
In the present study, it was found that treatment with
aqueous leaf extract of P. cineraria [Figure 1] at a
dose of 250
mg/kg body wt. was more effective
in producing antifertility effect when compared
to 150 mg/kg body wt. dose for 35 days. After 65
days, the antifertility effect was reversible. The
present study indicated that extract treatment did
not cause alterations in body weight of the treated
animals, suggesting that the treatment had no toxic
effect. Rajalakshmi, 1992, and Angela et al., 2010,
reported a reduction in the number of spermatozoa in
cauda epididymides in P. cineraria treated rat which
appeared to be due to the suppressive effect of P.
cineraria treatment on spermatogenesis in the sperm
number recovered to control levels after restoration
of spermatogenic activity following cessation of
treatment, while the alterations in sperm motility,
viability, and morphology might have resulted from
disturbances in epididymal function. A similar result
was observed in the present study on albino rats
treated with Andrographis paniculata (Sathiyaraj
et al., 2011). The treatment also caused significant
reduction in the weight of the seminal vesicle in
treated rats when compared to control animals.
Furthermore, the treatment also caused a marked
reduction in the level of the seminal vesicle in treated
rat, suggesting that P. cineraria treatment had an
adverse effect on the secretory function of the gland.
Table 1:
Antifertility
activity
of
Prosopis
cineraria
aqueous
leaf
extract
on
male
albino
rats:
Body
weight,
testis,
epididymis,
and
seminal
vesicle
weight
of
the
animal
Experiment
Body
weight
(g)
Reproductive
organs
weight
(g)
Total
sperm
count
m/ml
Motility
%
Abnormal
%
Testosterone
ng/ml
Initial
Final
Testis
Epididymis
Seminal
vesicle
Head
Tail
Group-I
control
210.16±1.47
212.66±4.71
1.45±0.55
1.14±0.41
0.83±0.11
8.20±1.69
95.83±1.56
2.01±0.62
2.02±0.62
4.92±0.55
Group-II
(150
mg/kg
body
wt.)
216.16±2.31
214.66±3.71
1.41±0.62*
1.01±0.63*
0.69±.28*
6.20±1.67*
58.50±2.50*
8.03±1.69*
8.03±1.51*
3.04±0.54*
Group-III
(250
mg/kg
body
wt.)
216.17±2.36
214.66±7.08
0.56±0.82*
0.68±0.34*
0.61±.34*
4.70±0.63*
58.50±1.69*
16.01±1.98*
14.01±1.69*
2.22±059*
Recovery
groups
212.51±3.26
218.18±1.44
1.43±0.27
0.75±0.13
0.72±0.18
10.08±0.466
95.81±0.67
3.35±0.28
3.30±0.30
4.71±0.66

5. Sathiyaraj, et al.: Antifertility effect: An male albino rat
IJPSCR/Jan-Mar-2021/Vol 1/Issue 1 20
In P. cineraria plant extract treated group, there
was a significant depletion in serum levels of
testosterone when compared to that of control
group. It is well known that testosterone plays a
major role in the maintenance of spermatogenesis
(O’Donell et al., 2006). Thus, it is probable
that P. cineraria-induced suppression of
spermatogenesis in rat testes in the present study
was due to deficiency of testosterone. At the
testicular level of testosterone secretion which is
responsible for diminished spermatogenesis and
hence, reduction in sperm counts (Mali et al.,
2002, Newman and Cragg, 2007; Sathiyaraj et al.,
2010; Thirumalai et al., 2012). The present study
also shows the reversible nature of P. cineraria
plant extract, since after the recovery period of
65 days, the animals became fertile and all the
altered parameters were restored to normal level
when compared to control. This study confirms
that the aqueous leaf extract of P. cineraria leaves
possesses anti-androgenic and anti-spermatogenic
activity and is likely to arrest spermatogenesis.
It also showed the reversible nature of the effect
of the extract after withdrawal of aqueous extract
treatment. An anti-androgenic effect is suggested
based on the regression and disintegration of
Leydig cells. Alterations of histological features
were more pronounced at high dose of 350 mg/
kg; disruption in seminiferous tubular arrangement
was observed with fewer Leydig cells in the present
study in male rat (Purohit and Dixit, 1991; Mishra
and Singh, 2009), had earlier reported alteration of
Leydig cell function in rats treated with aqueous
extracts of Prosopis cineraria. The nuclei became
pyknotic. Leydig cells revealed signs of atrophy.
Contrary to this, no morphological changes were
seen in the Sertoli cells.
CONCLUSION
This study reveals that the aqueous leaf extract
of P. cineraria has anti-fertility property when
administered alone. The toxic effects of the plant
on the testis may be due to direct deleterious
effects on the seminiferous tubules which are the
important testicular structures involved.
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